The use of retroviral pseudotypes as a vector for gene therapy is well documented. Recently, we have developed pseudotypes to determine neutralising antibody titres for highly pathogenic viruses. This removes the need for high containment facilities and allows small volumes of serum to be used. G-protein sequences from the rabies isolate CVS-11 and European bat lyssaviruses were cloned and co-expressed with lentiviral gag-pol and GFP or luciferase reporter genes. The pseudotypes infected a number of target cell lines and produced titres almost equivalent to VSV-G protein pseudotypes. Neutralisation assays using blinded sera and pseudotyped CVS-11 detected positive and negative samples with 100% specificity and sensitivity and correlated with the OIE FAVN (R2 = 0.89). It is possible to detect cross-neutralising antibodies to bat lyssaviruses pseudotypes.